Objective: To investigate the effects of Qiangjing Tablets (QJT) on sperm quality and the MAPK signaling pathway in the SD rat model of asthenospermia (AS).
Methods: A total of 100 male SD rats were randomly divided into five groups of equal number, blank control, AS model control, high-dose QJT, medium-dose QJT, and low-dose QJT. All the rats were intragastrically administered ORN at 200 mg/kg/d for establishment of the AS model except those in the blank control group, which were given 1% CMC sodium solution at 1 ml/100 g by gavage. Meanwhile the animals of the high-, medium-, and low-dose QJT groups were gavaged with QJT at 6700, 3300 and 1700 mg/kg/d, respectively, qd 6 days a week for 20 days. Then the testis issue and the apoptosis of the testicular cells were observed under the electron microscope, the expression of vimentin in the testis was determined with the immunohistochemical SP method, that of ERK1/2 detected by Western blot, and the concentration of TGF-β1 in the semen measured by ELISA.
Results: The AS model controls showed round nuclei of spermatocytes, homogeneously distributed chromatins, broken or lost mitochondria, and expanded rough endoplasmic reticulum in the testis tissue. In comparison, the rats of the high-, medium-, and low-dose QJT groups exhibited round nuclei of spermatocytes, homogeneously distributed chromatins, and well-structured mitochondria, rough endoplasmic reticulum and ribosome, which were all similar those of the blank controls. Compared with the blank controls, the AS model rats manifested significantly increased expressions of ERK1/2 (1.00 ± 0.00 vs 1.26 ± 0.10, P<0.01) and vimentin (0.16 ± 0.01 vs 0.17 ± 0.01, P<0.01) and apoptosis rate of cells in the testis tissue (［9.20 ± 3.07］ vs ［42.20 ± 9.17］ %, P<0.01), but decreased level of TGF-β1 in the semen (［627.67 ± 26.07］ vs ［566.73 ± 68.44］ ng/ml, P<0.05). In comparison with the model controls, the rats of the high- and medium- -dose QJT groups presented remarkably down-regulated expressions of ERK1/2 (1.26 ± 0.10 vs 1.14 ± 0.08, P<0.01; 1.26 ± 0.10 vs 1.18 ± 0.05, P<0.05) and vimentin (0.17 ± 0.01 vs 0.16 ± 0.01, P<0.01; 0.17 ± 0.01 vs 0.17 ± 0.09, P<0.05) and decreased rate of cell apoptosis (［42.20 ± 9.17］ vs ［21.60 ± 5.94］ %, P<0.01; ［42.20 ± 9.17］ vs ［33.95 ± 6.39］ %, P<0.05). The concentration of TGF-β1 in the semen was markedly lower in the high-dose QJT than in the AS model control group (［621.78 ± 30.80］ vs ［566.73 ± 68.44］ ng/ml, P < 0.05).
Conclusions: Qiangjing Tablets could improve semen quality in asthenospermia rats by acting against oxidative stress.
目的： 观察强精片对弱精子症SD大鼠精液质量及MAPK通路的影响。方法： 选取100只SD大鼠，采用单纯随机抽样方法分为空白对照组、模型组、强精片高剂量组、强精片中剂量组、强精片低剂量组，每组20只。除空白对照组外各组大鼠皆予奥硝唑（ORN）200 mg/(kg·d)灌胃造模，空白对照组大鼠1％羧甲基纤维素钠溶液1ml/100g灌胃，高剂量组、中剂量组、低剂量组均在灌胃ORN的同时，予不同剂量强精片：6 700 mg/(kg·d)、3 300 mg/(kg·d)、1 700 mg/(kg·d)，每周给药6 d，1次/d，连续20 d。实验结束后予电镜观察睾丸组织、细胞凋亡情况，采用S-P法测定睾丸波形蛋白表达，Western印迹测定大鼠睾丸ERK1/2表达，ELISA 法检测精液中TGF-β1表达量测定。结果： ①电镜结果显示模型组大鼠睾丸精母细胞，细胞核呈圆形，染色质分布均匀，胞浆内，线粒体的脊断裂或消失、严重肿胀，粗面内质网扩张；与模型组相比，高剂量组、中剂量组、低剂量组大鼠睾丸精母细胞，细胞核呈圆形，染色质分布均匀，胞浆内，线粒体，粗面内质网，核糖体等细胞器结构清晰，均与对照组类似。②空白对照组、模型组、高剂量组、中剂量组、低剂量组ERK1/2、波形蛋白相对表达量、睾丸组织细胞凋亡率（%）、精液TGF-β1表达量（ng/ml）分别为（1.00±0.00）、（1.26±0.10）、（1.14±0.08）、（1.18±0.05）、（1.19±0.19），（0.16±0.01）、（0.17±0.01）、（0.16±0.01）、（0.17±0.09）、（0.17±0.00），（9.20±3.07）、（42.20±9.17）、（21.60±5.94）、（33.95±6.39）、（40.85±5.61），（627.67±26.07）、（566.73±68.44）、（621.78±30.80）、（583.93±44.24）、（587.69±59.29）。模型组睾丸组织内ERK1/2、波形蛋白表达量高于对照组（P<0.01），模型组大鼠睾丸组织凋亡阳性细胞率较对照组明显升高（P≤0.01），而精液中TGF-β1表达量低于空白对照组（P≤0.05）；与模型组相比，高剂量组ERK1/2表达量、波形蛋白表达量均明显降低（P<0.01），中剂量组ERK1/2表达量、波形蛋白表达量均降低（P<0.05），高剂量组大鼠睾丸组织凋亡阳性细胞率明显降低（P≤0.01），中剂量组大鼠睾丸组织凋亡阳性细胞率降低（P≤0.05），而高剂量组精液中TGF-β1表达量明显高于模型组（P<0.05）。结论： 强精片可能是通过抗氧化应激改善弱精子症大鼠的精子质量。.
Keywords: MARK signaling pathway; Qiangjing Tablets; asthenospermia; rats.